RESUMO
Induction of terminal differentiation represents a potentially less toxic cancer therapy. Treatment of HO-1 human metastatic melanoma cells with IFN-ß plus mezerein (MEZ) promotes terminal differentiation with an irreversible loss of growth potential. During this process, the transcription factor FOXM1 is down-regulated potentially inhibiting transactivation of target genes including those involved in G(2)/M progression and cell proliferation. We investigated the mechanism of FOXM1 down-regulation and its physiological role in terminal differentiation. Genetic and pharmacological studies revealed that FOXM1 down-regulation was primarily caused by MEZ activation of PKCα and co-treatment with IFN-ß plus MEZ augmented the effect of PKCα. Promoter analysis with a mutated E-box on the FOXM1 promoter, and in vitro and in vivo binding assays confirm a direct role of c-Myc on FOXM1 expression. Reduction of c-Myc and overexpression of Mad1 by IFN-ß plus MEZ treatment should cause potent and persistent reduction of FOXM1 expression during terminal differentiation. Overexpression of FOXM1 restored expression of cell cycle-associated genes and increased the proportion of cells in the S phase. Our experiments support a model for terminal differentiation in which FOXM1 down-regulation via activation of PKCα followed by suppression of c-Myc expression, are causal events in promoting growth inhibition during terminal differentiation.
Assuntos
Fatores de Transcrição Forkhead/metabolismo , Regulação Neoplásica da Expressão Gênica/fisiologia , Melanoma/metabolismo , Antineoplásicos Fitogênicos/farmacologia , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/genética , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/metabolismo , Diferenciação Celular , Linhagem Celular Tumoral , Diterpenos/farmacologia , Proteína Forkhead Box M1 , Fatores de Transcrição Forkhead/genética , Humanos , Interferon beta/farmacologia , Melanoma/patologia , Regiões Promotoras Genéticas , Proteínas Proto-Oncogênicas c-myc/genética , Proteínas Proto-Oncogênicas c-myc/metabolismo , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo , Transdução de SinaisRESUMO
OBJECTIVE: To determine whether environmental surface contamination with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) occurred at a provincial hospital in Viet Nam that admitted patients with novel coronavirus disease 2019 (COVID-19) and at the regional reference laboratory responsible for confirmatory testing for SARS-CoV-2 in 2020. METHODS: Environmental samples were collected from patient and staff areas at the hospital and various operational and staff areas at the laboratory. Specimens from frequently touched surfaces in all rooms were collected using a moistened swab rubbed over a 25 cm2 area for each surface. The swabs were immediately transported to the laboratory for testing by real-time reverse transcription polymerase chain reaction (RT-PCR). Throat specimens were collected from staff at both locations and were also tested for SARS-CoV-2 using real-time RT-PCR. RESULTS: During the sampling period, the laboratory tested 6607 respiratory specimens for SARS-CoV-2 from patients within the region, and the hospital admitted 9 COVID-19 cases. Regular cleaning was conducted at both sites in accordance with infection prevention and control (IPC) practices. All 750 environmental samples (300 laboratory and 450 hospital) and 30 staff specimens were negative for SARS-CoV-2. DISCUSSION: IPC measures at the facilities may have contributed to the negative results from the environmental samples. Other possible explanations include sampling late in a patient's hospital stay when virus load was lower, having insufficient contact time with a surface or using insufficiently moist collection swabs. Further environmental sampling studies of SARS-CoV-2 should consider including testing for the environmental presence of viruses within laboratory settings, targeting the collection of samples to early in the course of a patient's illness and including sampling of confirmed positive control surfaces, while maintaining appropriate biosafety measures.
Assuntos
COVID-19 , SARS-CoV-2 , Hospitais , Humanos , Laboratórios , Vietnã/epidemiologiaRESUMO
@#As authorities braced for the arrival of the Omicron variant of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), infrastructure investments and government directives prompted action in central Viet Nam to establish capacity for genomic surveillance sequencing. From 17 November 2021 to 7 January 2022, the Pasteur Institute in Nha Trang sequenced 162 specimens from 98 150 confirmed SARS-CoV-2 cases in the region collected from 8 November to 31 December 2021. Of these, all 127 domestic cases were identified as the B.1.617.2 (Delta) variant, whereas 92% (32/35) of imported cases were identified as the B.1.1.529 (Omicron) variant, all among international flight passengers. Patients were successfully isolated, enabling health-care workers to prepare for additional cases. Most (78%) of the 32 Omicron cases were fully vaccinated, suggesting continued importance of public health and social measures to control the spread of new variants.
RESUMO
Defects in differentiation are frequently observed in cancer cells. By appropriate treatment specific tumor cell types can be induced to terminally differentiate. Metastatic HO-1 human melanoma cells treated with IFN-beta plus mezerein (MEZ) undergo irreversible growth arrest and terminal differentiation followed by apoptosis. In order to define the molecular changes associated with this process, changes in gene expression were analyzed by cDNA microarray hybridization and by semi-quantitative and quantitative RT-PCRs of representative 44 genes. The expression of 210 genes was changed more than two-fold at either 8 or 24 h post-treatment (166 up and 44 down). Major biological processes associated with the up-regulated genes were response to endogenous/exogenous stimuli (38%), cell proliferation (13%), cell death (16%) and development (30%). Approximately 34% of the down-regulated genes were associated with cell cycle, 9% in DNA replication and 11% in chromosome organization, respectively. Suppression of cell cycle associated genes appeared to directly correlate with growth arrest observed in the terminal differentiation process. Expression of Calpain 3 (CAPN3) variant 6 was suppressed by the combined treatment and maintained high in various melanoma cell lines. However, over-expression of the CAPN3 did not significantly affect growth kinetics and cell viability, suggesting that up-regulation of CAPN3 alone may not be a causative, but an associated change with melanoma development. This analysis provides further insights into the spectrum of up-regulated and the first detailed investigation of down-regulated gene changes associated with and potentially causative of induction of loss of proliferative capacity and terminal differentiation in human melanoma cells.